SLC4A11 and MFSD3 Gene Expression Changes in Deoxynivalenol Treated IPEC-J2 Cells
Yafei Xu1, Xiaolei Chen1, Luchen Yu1
1Key Laboratory for Animal Genetic, Breeding, Reproduction and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou, China.
Abstract:
Deoxynivalenol (DON) caused serious cytotoxicity for animal cells. However, genes involved in regulating DON toxicity and the underlying molecular mechanisms remain largely unknown. This study explored the role of SLC4A11 and MFSD3 in alleviating DON toxicity and analyzed the DNA methylation changes of these two genes. Viability and cell cycle analysis showed that DON exposure decreased the IPEC-J2 viability (P < 0.01), blocked the cell cycle in the G2/M phase (P < 0.01), and increased the rate of apoptosis (P < 0.05). Expression of the SLC4A11 and MFSD3 genes was significantly downregulated upon DON exposure (P < 0.01). Overexpression of SLC4A11 and MFSD3 can enhance the cell viability (P < 0.01). DNA methylation assays indicated that promoter methylation of SLC4A11 (mC-1 and mC-23) and MFSD3 (mC-1 and mC-12) were significantly higher compared with those in the controls and correlated negatively with mRNA expression (P < 0.05). Further analysis showed that mC-1 of SLC4A11 and MFSD3 was located in transcription factor binding sites for NF-1 and Sp1. Our findings revealed the novel biological functions of porcine SLC4A11 and MFSD3 genes in regulating the cytotoxic effects induced by DON, and may contribute to the detection of biomarkers and drug targets for predicting and eliminating the potential toxicity of DON.
Insights
Deoxynivalenol (DON) causes cell damage, but SLC4A11 and MFSD3 genes can alleviate its toxicity. Increased DNA methylation of these genes correlates with reduced expression and increased DON toxicity in animal cells.
Area of Science:
- Toxicology
- Molecular Biology
- Epigenetics
Background:
- Deoxynivalenol (DON) is a mycotoxin causing significant cytotoxicity in animal cells.
- The molecular mechanisms and regulatory genes involved in DON toxicity are not fully understood.
Purpose of the Study:
- To investigate the role of SLC4A11 and MFSD3 genes in mitigating DON-induced cytotoxicity.
- To analyze DNA methylation changes in SLC4A11 and MFSD3 and their correlation with DON toxicity.
Main Methods:
- Cell viability, cell cycle, and apoptosis assays were performed on IPEC-J2 cells exposed to DON.
- Gene expression levels of SLC4A11 and MFSD3 were analyzed.
- DNA methylation patterns in the promoter regions of SLC4A11 and MFSD3 were assessed using methylation assays.
- Bioinformatic analysis identified transcription factor binding sites.
Main Results:
- DON exposure decreased IPEC-J2 cell viability, arrested the cell cycle at G2/M phase, and increased apoptosis.
- Expression of SLC4A11 and MFSD3 was significantly downregulated by DON.
- Overexpression of SLC4A11 and MFSD3 enhanced cell viability.
- Increased promoter methylation of SLC4A11 and MFSD3 correlated negatively with their mRNA expression.
- Specific methylation sites (mC-1) in SLC4A11 and MFSD3 promoters were located within transcription factor binding sites (NF-1, Sp1).
Conclusions:
- Porcine SLC4A11 and MFSD3 play a crucial role in regulating DON-induced cytotoxicity.
- DNA methylation of SLC4A11 and MFSD3 promoter regions is involved in the modulation of DON toxicity.
- These genes represent potential biomarkers and therapeutic targets for predicting and mitigating DON toxicity.
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